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Molecular cloning,characterization and expression analysis of Sox9a and Foxl2 genes in half-smooth tongue sole(Cynoglossussemilaevis) 被引量:20
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作者 DONG Xiaoli CHEN Songlin +1 位作者 JI Xiangshan SHAO Changwei 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2011年第1期68-77,共10页
Two transcription factors, Sox9a and Foxl2 were cloned from half-smooth tongue sole (Cynoglossus semilaevis). Sox9a is a new duplication of C.semilaevis Sox9 gene. The complete cDNA of Sox9a gene was 1 842 bp long c... Two transcription factors, Sox9a and Foxl2 were cloned from half-smooth tongue sole (Cynoglossus semilaevis). Sox9a is a new duplication of C.semilaevis Sox9 gene. The complete cDNA of Sox9a gene was 1 842 bp long coding for 487 amino acids and Foxl2 gene was 1 817 bp coding for 308 amino acids. Sox9a was expressed higher in male brain, pituitary and gonad and Foxl2 were higher in female brain, pituitary and gonad. The expression of Sox9a gene in gonads of neo-males was higher than that of normal females. Sox9a and Foxl2 were expressed higher in gastrula stage than in other stages. In the period of sex differentiation, the expression of Sox9a was first going up and then going down and Foxl2 was higher expressed at 37 dph. The highest expressions of Sox9a and Foxl2 genes occurred in nine-month and 12-month old gonad tissues, respectively. Sox9a gene was considered to have inevitable links with sex reversal, sex differentiation and cell differentiation of embryos and formation of spermatogenic ceils. Foxl2 was considered to play a role in sex differentiation, cell differentiation of embryos but not to be necessary for sex determination and sex reversal. 展开更多
关键词 half-smooth tongue sole Cynoglossus semilaevis SOX9 FOXL2 real-time PCR sexreversal neo-male
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Isolation and Characterization of Pathogenic Listonella anguillarum of Diseased Half-Smooth Tongue Sole(Cynoglossus semilaevis Günther) 被引量:13
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作者 TANG Xiaoqian ZHOU Li ZHAN Wenbin 《Journal of Ocean University of China》 SCIE CAS 2008年第3期343-351,共9页
Studies were conducted to determine the cause of the acute mortality of half-smooth tongue sole Cynoglossus semilaevis Günther juveniles in a fish farm in Jimo, Shandong Province, China, in June 2006. Gross signs... Studies were conducted to determine the cause of the acute mortality of half-smooth tongue sole Cynoglossus semilaevis Günther juveniles in a fish farm in Jimo, Shandong Province, China, in June 2006. Gross signs of the diseased tongue sole included several petechiae and ecchymoses on the body and fin necrosis and hemorrhagic lesion at the base of the fin. Bacteria were isolated from kidney, liver and hemorrhagic lesions of the diseased tongue sole. Among 14 strains, SJ060621 was proved to be highly virulent to juvenile tongue sole with LD50 value of 〈1.0×10^5 colony forming units (CFU)mL^-1, while the remaining 13 were avirulent. Among the 16 antibiotics tested, SJ060621 was sensitive to gentamicin and nitrofurantoin. It was identified as Listonella anguillantm with conventional plate and tube tests in combination with API 20E analysis. 16S rRNA gene and partial HSP60 gene sequenceing analysis revealed that the strain was highly homologous with L. anguillarum. Examination of the infected musculature by electron microscopy indicated numerous bacteria and lots of macrophages containing phagocytosed bacteria. Histopathological investigations revealed severe necrotic degenerative changes in the infected organs. Indirect immunofluorescence assay (IFA) was employed to detect the location of occurrence of bacteria, and bacteria were found in aggregations in the inflammatory areas in musculature. 展开更多
关键词 half-smooth tongue sole Cynoglossus semilaevis Günther Listonella anguillarum 16S rRNA gene HSP60 gene HISTOPATHOLOGY IFA
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Analysis of SSRs Derived from an EST Library of Half-Smooth Tongue Sole Cynoglossus semilaevis 被引量:4
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作者 LIU Yunguo BAO Baolong +1 位作者 CHEN Songlin SUN Xiuqin 《Journal of Ocean University of China》 SCIE CAS 2008年第3期339-342,共4页
A complementary DNA(cDNA) library was constructed from half-smooth tongue sole spleen. A long-read expressed sequence tag(EST) database was generated,containing 3100 cDNA clones,of which 220 clones were fully sequence... A complementary DNA(cDNA) library was constructed from half-smooth tongue sole spleen. A long-read expressed sequence tag(EST) database was generated,containing 3100 cDNA clones,of which 220 clones were fully sequenced. A total of 1060 non-redundant simple sequence repeats(SSRs) were obtained from the cDNA library. An average of 5 kb sequence generates 1 SSR in the half-smooth tongue sole spleen cDNA library. The proportion of the SSR unit size was different in the cDNA library. The monomeric repeats(51.4%) are the most abundant class of SSR in the dataset. The dimeric,trimeric,tetrameric and hexameric re-peats are represented in decreasing proportions of 27.2%,16.0%,2.8% and 1.9%,respectively. The frequency of pentameric repeats was observed the least(only 0.7%) . Most of the monomeric and dimeric repeats are distributed in 3' and 5' un-translation region. If translation regions are considered merely,trimeric repeats are the highest,accounting for 57% of the total microsatellites. 展开更多
关键词 half-smooth tongue sole Cynoglossus semilaevis EST MICROSATELLITE
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Establishment and characterization of an astroglial cell line derived from the brain of half-smooth tongue sole(Cynoglossus semilaevis) 被引量:2
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作者 Tian-Zi WANG Ai SUN +3 位作者 Na WANG Zhong-Kai CUI Song-Lin CHEN Zhen-Xia SHA 《Zoological Research》 CAS CSCD 2015年第5期305-309,共5页
An astroglial cell line was established from the brain of half smooth tongue sole (Cynoglossus semilaevis) and was designated as CSAC. CSAC shows the morphological homogeneity of epithelial cells. The cell identity ... An astroglial cell line was established from the brain of half smooth tongue sole (Cynoglossus semilaevis) and was designated as CSAC. CSAC shows the morphological homogeneity of epithelial cells. The cell identity was tested by the presence of glial fibrillary acidic protein (GFAP), which was revealed by RT-PCR and immunofluorescence. The cell line was optimally maintained at 24 ℃ in minimum essential medium supplemented with HEPES, antibiotics, 20% fetal bovine serum, 2- Mercaptoethanol (2-Me) and basic fibroblast growth factor. Chromosome analysis revealed that the CSAC cells maintained a normal diploid chromosome number (2n=42). The fluorescent signals were observed in CSAC after the cells were transfected with green fluorescent protein (GFP) reporter plasmids. The CSAC cell line may serve as a valuable tool for studies on the potential functions of fish astroglial cells. 展开更多
关键词 half-smooth tongue sole Cynoglossussemilaevis Brain astroglial cell
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Phenotypic and genetic parameter estimation of juvenile growth and bottom color traits in half-smooth tongue sole,Cynoglossus semilaevis 被引量:2
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作者 LIU Feng 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2016年第10期83-87,共5页
Half-smooth tongue sole(Cynoglossus semilaevis) is a commercially important fish species. There were no reports about accurate genetic parameters of growth traits in C. semilaevis, despite efforts aimed at the cultu... Half-smooth tongue sole(Cynoglossus semilaevis) is a commercially important fish species. There were no reports about accurate genetic parameters of growth traits in C. semilaevis, despite efforts aimed at the culture of this species. Therefore, the main goal of this study was to estimate the parameters for the early growth traits(total length, body depth and body weight) of C. semilaevis based on the data obtained from 80 families. The trait of fish with pure white bottom or not(named as the trait of bottom color) was also analyzed. All the heritabilities estimated for the traits were medium, which varied from 0.210 to 0.362. Genetic correlations among growth traits were highly positive, varying from 0.913 to 0.959, indicating that selection of one of the three traits would result in correlated increase in the others. Also, genetic correlations between bottom color and growth traits were positive and varying from 0.241 to 0.353, suggesting that selection for the bottom color can be utilized to enhance the selection of growth traits. In addition, fish from the top performing 16 full-sib families were selected as broodstock, based on the above analysis and the comparison of family breeding value. This research provides an important basic material to implement selective breeding in C. semilaevis. 展开更多
关键词 half-smooth tongue sole Cynoglossus semilaevis growth trait bottom color genetic parameter family selection
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Application of AFLP markers for population genetic study on half-smooth tongue sole Cynoglossus semilaevis 被引量:2
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作者 刘云国 李俊峰 叶乃好 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2011年第3期609-615,共7页
The genetic diversity of wild and hatchery populations of half-smooth tongue sole Cynoglossus semilaevis, based on observation of amplified fragment length polymorphism (AFLP) was described. Two hundred individuals fr... The genetic diversity of wild and hatchery populations of half-smooth tongue sole Cynoglossus semilaevis, based on observation of amplified fragment length polymorphism (AFLP) was described. Two hundred individuals from four wild populations, Laizhou (LZ), Weihai (WH), Qingdao (QD), Rizhao (RZ), and one hatchery population, Mingbo (MB), were screened using eight different AFLP primer combinations. A total of 384 loci were screened in the five studied populations. 48.4%, 51.3%, 50.7%, 49.3% and 45.8% of these loci were polymorphic among the individuals tested in the LZ, WH, QD, RZ and MB populations, respectively. The number of polymorphic loci detected by single primer combinations ranged from 17 to 35. The average heterozygosity of the LZ, WH, QD, RZ and MB populations was 0.072, 0.093, 0.092, 0.090 and 0.063, respectively. The WH population showed the highest genetic diversity in terms of total number of AFLP bands, total number of polymorphic bands, average heterozygosity and percentage of low frequency (0-0.2) polymorphic loci among all the populations, while the LZ population was the lowest among the wild populations. Compared with the wild populations, the hatchery population showed a low genetic viability. 展开更多
关键词 half-smooth tongue sole Cynoglossus semilaevis AFLP marker genetic characteristics
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Histopathological Observation of Cynoglossus semilaevis Günther with Abdominal Dropsy in Industrial Aquaculture
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作者 Guangping GAO Guisheng GAO +7 位作者 Yanying ZHANG Zhixin JI Qinghui JIA Ting SU Qiumei SHI Hong-shengHAN Qingchun Song Jianxin ZHU 《Agricultural Biotechnology》 CAS 2016年第1期37-39,共3页
[ Objective] This study aimed to investigate and analyze the histopathological changes in various tissues and organs of Cynoglossus semilaevis Gunther with abdominal dropsy. [ Method] By HE staining, pathological tiss... [ Objective] This study aimed to investigate and analyze the histopathological changes in various tissues and organs of Cynoglossus semilaevis Gunther with abdominal dropsy. [ Method] By HE staining, pathological tissues of main organs of C. semilaevis Giinther with abdominal dropsy were observed. [ Result] According to the results of histopathological observation, C. semilaevis Gilnther with abdominal dropsy exhibited a large amount of ascites in the abdominal cavity; small intestinal mucosal epithelial cells were denatured and necrotic, and shed into the enteric cavity, with submucosal vascular congestion; the liver and pancreas tissues were denatured and necrotic, and the morphology of most liver cells disappeared. However, there was no remarkable pathological change in the heart, spleen and other organs. [ Conclusion] C. semilaevis Gtinther with abdominal dropsy exhibited serious pathological and histochemical lesions in the small intestine, liver and pancreas, causing metabolic disorders. 展开更多
关键词 half-smooth tongue sole (Cynoglossus semilaevis Giinther) Abdominal dropsy Histopathological observation
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Molecular characterization and expression analysis of Patched 1 gene in the half-smooth tongue sole(Cynoglossus semilaevis)
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作者 DONG Zhongdian QI Qian +4 位作者 ZHANG Ning SHAO Changwei ZHANG Liyan WEN Haishen CHEN Songlin 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2016年第6期19-28,共10页
The signaling molecule hedgehog(Hh) is essential for cellular signaling required for body pattern formation in embryonic and individual development in vertebrate and invertebrates.Patched 1(Ptc1),a receptor of Hh,... The signaling molecule hedgehog(Hh) is essential for cellular signaling required for body pattern formation in embryonic and individual development in vertebrate and invertebrates.Patched 1(Ptc1),a receptor of Hh,mediates the activity of Hh by binging the Hh ligands.In this study,the Ptc1 of half-smooth tongue sole Cynoglossus semilaevis(CsPtc1) was cloned and characterized.The full-length of CsPtc1 cDNA is 5 212 nucleotides(nt),and encodes a protein with 1 543 amino acid residues.CsPtc1 shared many features with Ptc1 of other telesot species.Real-time quantitative PCR revealed CsPtc1 mRNA expressed in brain,liver,heart,gill,intestines,spleen,gonad and kidney,with expression level in testis significantly higher than in ovary.In testis,hybridization signals were mainly detected in primary spermatocytes,secondary spermatocytes,and sertoli cells,while weak signals were found in oocytes.Correspondingly,the degree of methylation is higher in female than in male and pseudomale.Results indicate that CsPtc1 may be involved in Desert Hedgehog(DHH) maintenance of the male and pseudo-male germ line and spermatogenesis. 展开更多
关键词 half-smooth tongue sole male and female Patched 1 RACE QRT-PCR METHYLATION
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Characterization of the cyp19a1a gene from a BAC sequence in half-smooth tongue sole (Cynoglossus semilaevis) and analysis of its conservation among teleosts
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作者 SHAO Changwei LIU Geng +2 位作者 LIU Shanshan LIU Changlin CHEN Songlin 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2013年第8期35-43,共9页
The cyp19ala gene encodes an aromatase that plays a key role in sex differentiation of the gonad. The first bacterial artificial chromosome (BAC) sequence of half-smooth tongue sole (Cynoglossus sernilaeuis) conta... The cyp19ala gene encodes an aromatase that plays a key role in sex differentiation of the gonad. The first bacterial artificial chromosome (BAC) sequence of half-smooth tongue sole (Cynoglossus sernilaeuis) containing the intact cyp19ala gene was reported and the conservation and synteny of the cyp19ala gene among teleosts were analyzed in the study. The BAC is 107 367 bp in size, with an overall guanine-cytosine (GC) content of 43.44%, and contains 4.38% transposable elements. Nine genes were predicted, including seven functional genes and two hypothetical genes. The cyp19ala gene of all tested teleosts has nine exons and eight introns, and potential binding sites flanking the transcriptional start site are conserved. The ex- pression pattern among teleosts is also similar during ovarian differentiation. Synteny analysis revealed a conserved gene cluster PKH4B-SL9A5-FHOD3-CEBPG-CEBPA among teleosts. These findings suggest that, among teleosts, cyp19ala genes not only have similar genomic structures, but also have conserved function- s. The genomic environment of cyp19ala in tongue sole is not universal in teleosts, reflecting the particular evolution of tongue sole cyp19ala after it diverged from the other teleosts. 展开更多
关键词 BAC cypl9ala CONSERVATION gene synteny half-smooth tongue sole
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Construction of a microsatellite-based genetic linkage map for half-smooth tongue sole Cynoglossus semilaevis 被引量:1
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作者 Wentao SONG Guidong MIAO +5 位作者 Yongwei ZHAO Yuze NIU Renyi PANG Xiaolin LIAO Changwei SHAO Songlin CHEN 《Current Zoology》 SCIE CAS CSCD 2013年第1期99-108,共10页
The half-smooth tongue sole Cynoglossus semilaevis is an important cultured marine fish and a promising model fish for the study of sex determination. Sex-specific genetic linkage maps of half-smooth tongue sole were ... The half-smooth tongue sole Cynoglossus semilaevis is an important cultured marine fish and a promising model fish for the study of sex determination. Sex-specific genetic linkage maps of half-smooth tongue sole were developed with 567 mark- ers (565 microsatellite markers and two SCAR markers). The parents and F1 progeny (92 individuals) were used as segregating populations. The female map was composed of 480 markers in 21 linkage groups, covering a total of 1388.1 cM, with an average interval 3.06 cM between markers. The male map consisted of 417 markers in 21 linkage groups, spanning 1480.9 cM, with an average interval of 3.75 cM. The female and male maps had 474 and 416 unique positions, respectively. The genome length of half-smooth tongue sole was estimated to be 1522.9 cM for females and 1649.1cM for males. Based on estimations of map length, the female and male maps covered 91.1% and 89.8% of the genome, respectively. Furthermore, two female-specific SCAR mark- ers, f-382 and f-783, were mapped on LG15f (linkage group 15 in female maps). The present study presents a mid-density genetic linkage map for half-smooth tongue sole. These improved genetic linkage maps may facilitate systematic genome searches to identify quantitative trait loci (QTL), such as disease resistance, growth and sex-related traits, and are very useful for marker-assisted selection breeding programs for economically important traits in half-smooth tongue sole [Current Zoology 59 (1): 99-108, 2013]. 展开更多
关键词 half-smooth tongue sole Cynoglossus semilaevis MICROSATELLITE Genetic linkage map MAS
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Development of 57 Novel Polymorphic Microsatellite Markers in Half-Smooth Tongue Sole(Cynoglossus semilaevis)
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作者 MIAO Guidong XU Ying +3 位作者 WANG Di CHEN Songlin FAN Tingjun TIAN Yongsheng 《Journal of Ocean University of China》 SCIE CAS 2011年第3期241-245,共5页
Half-smooth tongue sole (Cynoglossus semilaevis) is a promising species for aquaculture in China.The wild population of C. semilaevis is under threat from environmental factors. Microsatellite markers are very suitabl... Half-smooth tongue sole (Cynoglossus semilaevis) is a promising species for aquaculture in China.The wild population of C. semilaevis is under threat from environmental factors. Microsatellite markers are very suitable for assessing genetic diversity. Four microsatellite-enriched libraries of half smooth tongue sole (Cynoglossus semilaevis) were constructed,from which 57 polymorphic microsatellites were isolated and characterized.The polymorphism of these microsatellites was assessed by genotyping in 30 individual fish.The number of alleles ranged from 2 to 11, with an average of 4.614 alleles per locus.The values of observed and expected heterozygosities ranged from 0.1000 to 1.0000 and from 0.0966 to 0.8847 respectively. Polymorphism information content (PIC) ranged from 0.0905 to 0.862.These markers would be useful for population structure assessment,genetic linkage map construction and parentage analysis for this species. 展开更多
关键词 Cynoglossus semilaevis MICROSATELLITE polymorphic marker half-smooth tongue sole
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半滑舌鳎病原鳗利斯顿氏菌表型及分子特征研究 被引量:34
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作者 张晓君 秦国民 +3 位作者 阎斌伦 徐静 毕可然 秦蕾 《海洋学报》 CAS CSCD 北大核心 2009年第5期112-122,共11页
2008年12月,江苏赣榆某渔场养殖半滑舌鳎出现大量死亡,症状主要表现为:头部、鳃盖及鳍基出血,尾鳍腐烂,腹腔膨胀并积有大量腹水,部分病鱼肠管脱出肛外。从病鱼肝脏、腹水中分离出大量优势生长的细菌,人工感染试验证明其对半滑舌鳎有较... 2008年12月,江苏赣榆某渔场养殖半滑舌鳎出现大量死亡,症状主要表现为:头部、鳃盖及鳍基出血,尾鳍腐烂,腹腔膨胀并积有大量腹水,部分病鱼肠管脱出肛外。从病鱼肝脏、腹水中分离出大量优势生长的细菌,人工感染试验证明其对半滑舌鳎有较强的致病性。对3株分离菌进行了形态特征、理化特性、胞外酶及溶血素活性等表型生物学性状检验;测定了代表菌株的16SrRNA和gyrB基因序列,分析了16 S rRNA和gyrB两种基因序列的同源性,并构建了系统发生树,比较了两种基因对分离菌的鉴定能力;结果表明gyrB基因用于细菌种间鉴定更具优越性;基于16S rRNA和gyrB基因的系统发育学分析表明分离菌与鳗利斯顿氏菌具有高同源性,根据分离菌的表型及分子特征,判定分离菌为利斯顿氏菌属(Listonella MacDonell and Colwell 1986)的鳗利斯顿氏菌[Listonella anguillarum (Bergeman 1909) MacDonell and Colwell 1986]。胞外酶活性及溶血活性检测表明3株分离菌均具有淀粉酶、蛋白酶、卵磷脂酶等胞外酶活性,在含7%家兔脱纤血液营养琼脂培养基上呈β型溶血,PCR检测3株供试菌均可扩增出大小约493 bp的溶血素基因和大小约248 bp的金属蛋白酶基因。分离菌的耐药谱测定结果显示,对供试49种抗菌药物中的青霉素G等13种药物耐药,对羧苄青霉素等6种药物存在敏感与耐药的株间差异。 展开更多
关键词 半滑舌鳎(Cynoglossusse milaevis Günther) 鳗利斯顿氏菌(Listonella anguillarum) 16S RRNA基因 gyrB基因
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石斑鱼和半滑舌鳎封闭循环水养殖系统的构建与运用 被引量:29
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作者 辛乃宏 于学权 +3 位作者 吕志敏 杨永海 朋礼全 张树森 《渔业现代化》 北大核心 2009年第3期21-25,40,共6页
介绍适合于点带石斑鱼(Epinephelus malabaricus)和半滑舌鳎(Cynoglossus semilaevlis Günther)的循环水养殖系统的工艺流程、生物调控及管理措施。通过在系统中采用弧形筛和蛋白分离器组合装置替代微滤机、添加比表面积大的塑料... 介绍适合于点带石斑鱼(Epinephelus malabaricus)和半滑舌鳎(Cynoglossus semilaevlis Günther)的循环水养殖系统的工艺流程、生物调控及管理措施。通过在系统中采用弧形筛和蛋白分离器组合装置替代微滤机、添加比表面积大的塑料片状滤料、增加臭氧和紫外线2级灭菌装置等手段,并且针对循环水养殖的特点制定系统的生物调控和管理规范,使该养殖系统石斑鱼和半滑舌鳎的养殖承载量分别达到30 kg/m2和20 kg/m2;养殖1 kg海水鱼的电耗为7.54 kW,并能达到无公害水产品的质量要求。 展开更多
关键词 石斑鱼 半滑舌鳎 循环水养殖 生物调控
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基于高通量测序的池塘养殖半滑舌鳎消化道菌群的结构特征分析 被引量:23
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作者 张正 李彬 +3 位作者 王印庚 廖梅杰 王岚 荣小军 《水生生物学报》 CAS CSCD 北大核心 2015年第1期38-45,共8页
运用高通量测序方法研究了池塘养殖半滑舌鳎消化道和池水、池塘底泥、颗粒饵料中的微生物菌群结构,并分析了它们之间的相关性。实验结果证实,养殖池塘池水、底泥和饵料中的细菌种类多样性要远高于半滑舌鳎消化道中的细菌多样性,但消化... 运用高通量测序方法研究了池塘养殖半滑舌鳎消化道和池水、池塘底泥、颗粒饵料中的微生物菌群结构,并分析了它们之间的相关性。实验结果证实,养殖池塘池水、底泥和饵料中的细菌种类多样性要远高于半滑舌鳎消化道中的细菌多样性,但消化道中的菌群结构与池水和池塘底泥中的细菌没有相关性,仅在一定程度上受到颗粒饵料中菌群的影响。而半滑舌鳎消化道不同部位的细菌种类多样性存在一定的差异,但优势菌群的组成并没有明显的结构差异,优势菌种的构成和相对数量比例非常相似,并且大部分的优势菌种在NCBI数据库中没有同源菌株。研究说明半滑舌鳎消化道菌群这一微生态系统的结构相对独立,在池塘养殖这一阶段几乎不受养殖环境中菌群的影响,而且定植肠道中的一些优势细菌种类可能具有宿主特异性。 展开更多
关键词 半滑舌鳎 池塘养殖 菌群结构 高通量测序
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程序化设计的简并引物克隆半滑舌鳎CYP17基因 被引量:21
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作者 陈彩芳 温海深 +1 位作者 何峰 董双林 《中国海洋大学学报(自然科学版)》 CAS CSCD 北大核心 2009年第6期1213-1218,共6页
采用1种新的简并引物设计方法——CODEHOP法来克隆半滑舌鳎CYP17基因。该法结合NCBI、Blastp、BlockMaker、CODEHOP等生物信息学资源和DNAMAN、Oligo6.0等生物软件,程序性设计半滑舌鳎CYP17基因的CODEHOP引物,从半滑舌鳎卵巢组织中成功... 采用1种新的简并引物设计方法——CODEHOP法来克隆半滑舌鳎CYP17基因。该法结合NCBI、Blastp、BlockMaker、CODEHOP等生物信息学资源和DNAMAN、Oligo6.0等生物软件,程序性设计半滑舌鳎CYP17基因的CODEHOP引物,从半滑舌鳎卵巢组织中成功克隆了CYP17基因,该基因片段与其它物种的CYP17家族具有较高的同源性,经鉴定,该基因属于整个脊椎动物的CYP17基因群,尤其是鱼类亚群。实验结果表明,程序性设计的简并引物较传统简并引物特异性更高,假阳性率少,更有利于实验成功。 展开更多
关键词 半滑舌鳎 简并引物 CYP17
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半滑舌鳎脑芳香化酶基因cDNA克隆及表达分析 被引量:15
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作者 邓思平 陈松林 +2 位作者 刘本伟 徐建勇 田永胜 《Zoological Research》 CAS CSCD 北大核心 2008年第1期17-24,共8页
为研究脑型芳香化酶(P450aromB)在半滑舌鳎性别分化中的作用,采用同源克隆策略,从半滑舌鳎脑分离了2184bp长的脑型芳香化酶的全长cDNA,该基因编码498个氨基酸。氨基酸序列和系统发育分析表明,P450aromB属于脑型P450arom,P450aromB的氨... 为研究脑型芳香化酶(P450aromB)在半滑舌鳎性别分化中的作用,采用同源克隆策略,从半滑舌鳎脑分离了2184bp长的脑型芳香化酶的全长cDNA,该基因编码498个氨基酸。氨基酸序列和系统发育分析表明,P450aromB属于脑型P450arom,P450aromB的氨基酸序列与其他鱼类脑型P450arom的同源性较高(48.3%-66.1%),与性腺型P450arom的同源性较低(34.2%-49.9%),与自身的性腺型芳香化酶同源性为45.1%。RT-PCR分析表明:P450aromB mRNA的表达具有明显组织特异性,P450aromB只在性腺、脑、鳃和皮肤中表达,且脑中表达量远高于性腺,而在雌雄鱼的其他组织中都不表达。经过甲基睾酮浸浴处理和高温诱导半滑舌鳎由雌性性反转为雄性后,脑中P450aromB的表达量降低,这些结果表明P450aromB参与了半滑舌鳎的性腺分化和性别决定过程。 展开更多
关键词 半滑舌鳎 芳香化酶 CDNA克隆 表达
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半滑舌鳎精子发生和精子形成的超微结构 被引量:15
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作者 康现江 梁春光 +4 位作者 郭明申 葛少钦 穆淑梅 苏文清 刘学会 《动物学报》 SCIE CAS CSCD 北大核心 2008年第2期356-365,共10页
用电子显微镜对半滑舌鳎(Cynoglossus semilaevis)精子发生的过程及精子的超微结构进行了观察。半滑舌鳎精巢属于小叶型,精小叶由各期生精细胞和支持细胞构成。半滑舌鳎的精子发生经历了初级精原细胞、次级精原细胞、初级精母细胞、次... 用电子显微镜对半滑舌鳎(Cynoglossus semilaevis)精子发生的过程及精子的超微结构进行了观察。半滑舌鳎精巢属于小叶型,精小叶由各期生精细胞和支持细胞构成。半滑舌鳎的精子发生经历了初级精原细胞、次级精原细胞、初级精母细胞、次级精母细胞和精子细胞,再经过精子形成过程发育成为精子。初级精母细胞成熟分裂的前期Ⅰ,同源染色体经历了联会复合体形成和解聚的变化。在精子形成的过程中,精细胞大致经历了核质浓缩、线粒体迁移及鞭毛的发生等过程。核质浓缩时,精细胞核内位于植入窝周围的染色质首先由细颗粒状浓缩成粗大颗粒状,然后细胞核其他部位的染色质也逐渐浓缩成粗大颗粒状。这些已浓缩成粗大颗粒状的染色质再进一步浓缩为电子密度高的均匀状物质。随着核质的浓缩,核外膜与核内膜之间的间隙增大形成核膜间隙,核内一些没有参与染色质浓缩的物质通过出芽形成囊泡,先排入核膜间隙,然后再外排到细胞质中。核浓缩过程中细胞核的体积和表面积都大大缩小;鞭毛的形成与细胞核的浓缩是同步进行的,当一对中心粒移近细胞核时,核膜凹陷形成植入窝,其周围染色质浓缩的同时,远端中心粒(基体)逐渐向后产生轴丝。成熟精子无顶体,头细长,主要为核占据,核凹窝发达,线粒体4-5个环绕在鞭毛基部形成袖套,尾细长,具侧鳍,尾部轴丝为"9+2"结构。 展开更多
关键词 半滑舌鳎 精巢 精子发生 超微结构 联会复合体
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半滑舌鳎雌性特异AFLP标记CseF783的克隆及其在遗传性别鉴定中的应用 被引量:15
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作者 马洪雨 陈松林 +3 位作者 李静 田永胜 季相山 张立敬 《遗传》 CAS CSCD 北大核心 2009年第1期88-94,共7页
半滑舌鳎性别控制和全雌育种等研究领域中迫切需要一种能够快速鉴定鱼类个体遗传性别的有效方法。文章采用AFLP技术,利用选择性引物组合(E-ACT/M-CAA)从半滑舌鳎中筛选到一条雌性特异的AFLP标记。对该标记进行二次PCR扩增、琼脂糖凝胶... 半滑舌鳎性别控制和全雌育种等研究领域中迫切需要一种能够快速鉴定鱼类个体遗传性别的有效方法。文章采用AFLP技术,利用选择性引物组合(E-ACT/M-CAA)从半滑舌鳎中筛选到一条雌性特异的AFLP标记。对该标记进行二次PCR扩增、琼脂糖凝胶回收、克隆、测序。分析表明,序列全长为791 bp,与GenBank中的序列无同源性。以该雌性特异AFLP标记DNA序列为模板,设计了一对特异的PCR引物,成功地将其转化为SCAR(Sequence characterized amplified regions)标记,并在100尾已知性别的半滑舌鳎个体(雌雄各50尾)中进行验证,结果表明,该SCAR标记在所有雌性个体中均扩增得到一条长度为324 bp的DNA条带,而在49尾雄性个体中均扩增不到该DNA条带(有1尾雄性个体例外),证明该SCAR标记是雌性特异的,并可用于半滑舌鳎个体遗传性别鉴定。随后,利用该SCAR标记检测了3日龄半滑舌鳎幼苗,结果表明,雌性个体比例为41.7%。 展开更多
关键词 半滑舌鳎 雌性特异AFLP标记 SCAR标记 遗传性别 鉴定
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半滑舌鳎肝脏细胞系的建立与鉴定 被引量:11
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作者 任国诚 陈松林 沙珍霞 《高技术通讯》 CAS CSCD 北大核心 2008年第6期657-660,共4页
以半滑舌鳎肝脏组织为材料,探索了组织细胞分离培养条件,建立了半滑舌鳎组织细胞培养技术及半滑舌鳎肝脏细胞系(HTLC)。该细胞系的培养基是添加了抗生素、胎牛血清(FBS)、花鲈血清(SPS)、成纤维生长因子(bFGF)的 MEM。HTLC 形态呈纤维状... 以半滑舌鳎肝脏组织为材料,探索了组织细胞分离培养条件,建立了半滑舌鳎组织细胞培养技术及半滑舌鳎肝脏细胞系(HTLC)。该细胞系的培养基是添加了抗生素、胎牛血清(FBS)、花鲈血清(SPS)、成纤维生长因子(bFGF)的 MEM。HTLC 形态呈纤维状,在培养基中生长迅速,经200多天的培养,成功传代30多代。检测了温度、FBS 浓度、bFGF对其生长的影响,结果表明,在24~30℃生长良好,但当温度低于12℃时细胞生长速度明显减慢;在一定浓度范围内,其生长速度随血清浓度的升高而增快,血清浓度过高或过低对其生长不利;在培养基中添加 bFGF 可以使细胞生长速度显著提高;其二倍体核型为2n=21t。将绿色荧光蛋白(GFP)报告基因通过脂质体介导的方法转入 HTLC 中并成功地获得了表达,转化率为20%左右。该细胞系的建立为研究鱼类病毒学、免疫学、遗传学、功能基因组学提供了很好的实验材料。 展开更多
关键词 半滑舌鳎 肝脏细胞系 核型 GFP报告基因
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半滑舌鳎人工催产技术研究 被引量:10
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作者 杨景峰 陈松林 +3 位作者 翟介明 田永胜 苏鹏志 孙丽娟 《内蒙古民族大学学报(自然科学版)》 2010年第2期185-187,190,共4页
为了提高半滑舌鳎(Cynoglossus semilaevis)苗种生产的效率,获得批量合格的受精卵,采取控温控光和营养强化培育等措施,使亲鱼性腺全部发育,进行人工催产和人工授精的研究.结果表明多种激素及其激素组合均能有效的对半滑舌鳎进行催产,其... 为了提高半滑舌鳎(Cynoglossus semilaevis)苗种生产的效率,获得批量合格的受精卵,采取控温控光和营养强化培育等措施,使亲鱼性腺全部发育,进行人工催产和人工授精的研究.结果表明多种激素及其激素组合均能有效的对半滑舌鳎进行催产,其中以LRH-A3+DOM的效果较好.在5种催产方案中23℃的效应时间为35-52h,各方案之间差异不显著.人工催产卵的卵径为1.02±0.13mm,明显低于自然产卵的卵径,但受精率、出卵率和孵化率和对照组差异不明显.统计其中10条鱼的催产情况;总浮卵率89.4%,总受精率65.3%,总孵化率为44.8%.共收孵化前卵1060ml,孵化鱼苗约83万尾. 展开更多
关键词 半滑舌鳎 人工催产 激素 效应时间
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